This application is a national stage of PCT/EP99/08269 filed Oct. 29, 1999, and based upon Argentina Application No. P980105446 filed Oct. 30, 1998, under the International Convention.
The present invention relates to novel uses of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid or any of its soluble salts or any of its hydrates.
1. Field of the Invention
The compound 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid, or any of its soluble salts or any of its hydrates, belongs to the group of bisphosphonates which are compounds containing two phosphonate groups bound to a carbon and two additional groups R1 and R2, respectively, which bind strongly to calcium crystals, inhibit their growth, suppress bone resorption and are being used in the treatment of a variety of disorders of bone metabolism.
2. Summary of the Invention
1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid is the amino-substituted form of olpadronate (OPD) and is hereafter sometimes referred to as NH2-OPD.
Bisphosphonates are recognized potent inhibitors of bone resorption and are successfully used in the treatment of common bone disorders such as osteoporosis and tumoral bone diseases, amongst others. Structurally, bisphosphonates are derivatives of methylene bisphosphonate in which the two hydrogens on the geminal carbon atom are replaced by two groups, namely R1 and R2, which usually are not identical. In many cases the R1 moiety is a hydroxyl group, because of its ability to form a tridentate structure together with the two phosphonate groups to bind bone material.
Extensive studies have been carried out on the effects of bisphosphonates on bone cells, in particular those related to their anti-resorptive potency. The molecular mechanisms involved in such actions, however, are not clear. The anti-resorptive activity and its utility in the treatment of osteoporoses and related bone diseases have, for example, been reported and described in the following patents: WO 94/00129 to Frances M. et al., WO 93/11786 to Gueddes A. et al., WO 92/14474 to McOskar et al., U.S. Pat. No. 4,942,157 to Gall R. et al., and in U.S. Pat. No. 3,962,432 to Schmidt-Dunker M.
Although it appears that the nature of the R2 moiety of bisphosphonates determines their anti-resorptive activity, replacement of the hydroxyl group in R1 by an NH2 group in some bisphosphonates (e.g. olpadronate and pamidronate) markedly reduces their anti-resorptive activity. This decrease in anti-resorptive efficacy of the amino derivatives is thought to be due to substitution-dependent changes in the cellular effects of the compounds, since their affinity for bone mineral is not significantly affected. This lack of anti-resorptive activity is, for example, disclosed in vanBeek et al., 1996, Journal of Bone and Mineral Research, vol. 11, no. 10, pp. 1492-1497, and international patent application PCT/EP96/02981, the latter disclosing the use of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid for the treatment of osteoporoses, arthritis and periodontal diseases.
The structure and stability of skeletal bones isxe2x80x94to a high degreexe2x80x94influenced and determined by an equilibrium between the activities of osteoblasts and osteoclasts, i.e. the cells responsible for mineralization and resorption of bone structure, respectively. Therefore, bones are subject to a continuous restructuring process involving anabolic and catabolic reactions. In human beings, as of the age of 30, the catabolic processes prevail such that there will be a net loss in total bone substance naturally. Consequently, the rate at which the renewal of the bone tissue takes place will be lowered with increasing age.
Object of the present invention is to provide means for prevention of the onset of clinically pathological bone-conditions.
Another object of the present invention is to avoid a predisposition to bone disfunctions or bone diseases.
Yet a further object of the present invention is to optimize the physiological bone functions in patients.
The present invention in its various aspects and embodiments disclosed hereafter solves all these objectives and provides further advantages.
The present invention is related to novel uses of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid and/or any of its soluble salts and/or any of its hydrates. For the purposes of the following specification, it is intended that the term 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid also encompasses any of its soluble salts and/or any of its hydrates.
Briefly stated, the present invention provides novel uses of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid, medicaments comprising 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid and screening methods involving 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid.
Within one aspect of the present invention, there is provided the use of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid for the manufacture of a medicament for selective modulation of osteoblasts. Furthermore, there is provided the use of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid for the manufacture of a medicament for the maintenance of a healthy bone structure.
There is also provided the use of b 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid for the manufacture of a medicament for the prevention of osteopathies in healthy patients.
The invention also provides for the use of 1-amino-3-(N,N-diethylamino)-propylidene-1,1-bisphosphonic acid for the manufacture of a medicament for the treatment of patients who have recently undergone treatment with corticosteroids.
Additionally there is provided the use of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid for the manufacture of a medicament for post-treatment of osteopathies wherein an anti-resorptive activity is not desired.
The invention also provides for the use of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid for the manufacture of a medicament for the treatment of children having an osteopathy.
In another aspect of the invention, there is provided the use of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid for the manufacture of a medicament for the stimulation of both signalling cascades and reaction mechanisms mediating the action of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid which can be blocked by Ca2+-channel blockers.
In yet another aspect of the present invention, there is provided the use of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid for the manufacture of a medicament for the mobilization of Ca2+-ions from IP3 sensitive stores.
In yet another aspect of the invention, there is provided a method for screening for Ca2+-channel blockers comprising the steps:
treatment of cells having Ca2+-channels with a putative Ca2+-channel blocker;
contacting the cells with 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid;
measuring a response as a result of the contacting step.
It is envisioned that the treatment-step and the contacting-step occur sequentially with the treatment-step preceding the contacting-step or vice versa, or that they occur simultaneously.
In another aspect, the invention also provides a method for screening for functional analogues of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid, comprising the steps:
treatment of cells having Ca2+-channels with Ca2+-channel blockers;
contacting the cells with the putative functional analogue which, in the absence of any Ca2+-channel blockers, is known to cause a Ca2+-ion influx into the cells;
measuring a response as a result of the contacting step.
It is envisioned that the treatment-step and the contacting-step occur sequentially with the treatment-step preceding the contacting-step or vice versa, or that they occur simultaneously.
In the present invention there is also disclosed a medicament comprising 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid and at least one physiologically acceptable carrier, for the selective modulation of the osteoblasts, for the maintenance of a healthy bone structure, for the prevention of osteopathies or for the post-treatment of osteopathies where an anti-resorptive activity is not desired.
In one aspect of the present invention it provides a medicament comprising 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid, which is to be applied in healthy patients or in patients at or above 40 years of age.
The present invention also discloses a method for the selective modulation of osteoblasts and/or for the maintenance of a healthy bone structure and/or for the prevention of osteopathies in healthy patients and/or for the treatment of patients who have recently undergone treatment with corticosteroids, and/or for post-treatment of osteopathies where an anti-resorptive activity is not desired, and/or for the treatment of children having an osteopathy and/or for the stimulation of those signaling cascades and reaction mechanisms mediating the action of 1-amino-3-(N,N-diethylamino)-propylidene-1,1-bisphosphonic acid or any of its soluble salts or any of its hydrates, which can be blocked by Ca2+-channel blockers, and/or for the mobilization of Ca2+ ions from IP3-sensitive stores, comprising
administering 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid or any of its soluble salts or any of its hydrates alone or in combination with a pharmaceutical carrier to a patient, the 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid or any of its soluble salts or any of its hydrates being administered in doses of 0.1 to 1000 mg/oral application or 0.02 to 200 mg/parenteral application.
It is preferred that the term xe2x80x9cselective modulation of osteoblastsxe2x80x9d comprises a stimulation of the cellular activities of the osteoblasts, more preferably an influence on the Ca2+-homeostasis of the osteoblasts. A preferred embodiment of xe2x80x9cselective modulationxe2x80x9d comprises a transient increase of the Ca2+-levels in the osteoblasts. A selective modulation of osteoblasts may also encompass and manifest itself in the synthesis of osteocalcine and/or osteopontin, osteonectin, calprotectin, fibronectin, matrix Gla protein and/or bone sialoprotein.
It is intended in the present invention that the term xe2x80x9cmaintenance of a healthy bone structurexe2x80x9d also encompass a prevention of clinically pathological conditions or diseases.
Within one aspect of the present invention it is preferred that where 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid is used for the manufacture of a medicament for the maintenance of healthy bone structure, that the medicament is applied in healthy patients, more preferably patients without osteopathies. The term xe2x80x9chealthy bone structurexe2x80x9d is meant to include any bone structure characterized by a balanced equilibrium between catabolic and anabolic processes resulting in a structure capable of withstanding mechanical stress that occurs as a result of the organism""s activities, as well as a structure that is capable of remodelling itself by aforementioned equilibrium and repairing itself by the removal of weakened or worn-out parts and the build-up of strong parts, thereby avoiding microcracks and parts prone to fracture.
It has been found that, especially with older people, the bones have a less strong structure because of their reduced rate of renewal. Furthermore, older people may get weaker, more fragile bones due to sedentary lifestyle, muscular weakness and insufficient nutrition. As a result the affected bones, in fact, comprise xe2x80x9cfitxe2x80x9d mineralized structures and therefore become brittle and unstable which very often is accompanied by a tendency for fractures. These xe2x80x9cunfitxe2x80x9d structures have not manifested themselves yet as clinically pathological conditions but are less than ideal. They can also be foundxe2x80x94apart from in people at or above the age of 40 yearsxe2x80x94in people recently treated with corticosteroids or in people recently treated with anti-osteoporotic agents such as fluorine and common bisphosphonates such as etidronate and chlodronate.
The term xe2x80x9cpatientxe2x80x9d is meant to encompass human beings as well as any vertebrate animal, for example cats, dogs, cows, horses and other domestic animals.
The term xe2x80x9costeopathyxe2x80x9d designates any pathological condition of the bone resulting in a weakened or irregular or abnormal bone structure. Within the present invention it is used synonymously with xe2x80x9cbone diseasexe2x80x9d. Preferably the term xe2x80x9costeopathyxe2x80x9d refers to a clinically pathological condition selected from the group comprising osteoporosis, Paget""s disease, arthritis, periodontal osteopenia, adolescent scoliosis, fracture, disuse osteopenia, post-transplant osteopenia, hyper-parathyroidism-associated osteopenia, drug-induced osteopenia, nutritional osteopenia, metabolic bone disease, osteopenia of prematurity and ossification disorder.
Where the term xe2x80x9cpatientxe2x80x9d is referring to a human being, in one embodiment it is preferred that the medicament is applied in human beings at or above the age of 40 years, in another embodiment it is preferred that the medicament is applied in children. Preferably, the term xe2x80x9cchildrenxe2x80x9d is meant to encompass individuals who are from 0 to 16 years old.
Where 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid is used for the manufacture of a medicament for the prevention of osteopathies in healthy patients or for the manufacture of a medicament for the treatment of children having an osteopathy it is preferred that the osteopathy is selected from the group comprising osteoporosis, Paget""s disease, arthritis, periodontal osteopenia, adolescent scoliosis, fracture, disuse osteopenia, post-transplant osteopenia, hyper-parathyroidism-associated osteopenia, drug-induced osteopenia, nutritional osteopenia, metabolic bone disease, osteopenia of prematurity and ossification disorder.
In one embodiment it is preferred that 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acidxe2x80x94after the application of the medicament in a patientxe2x80x94will be present at extracellular concentrations in a range between 10xe2x88x926M and 10xe2x88x9210M, more preferably 10xe2x88x927M and 10xe2x88x929M, most preferably at an extracellular concentration of about 10xe2x88x928M.
In the embodiment where 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid is used for the manufacture of a medicament for the stimulation of those signaling cascades and reaction mechanisms mediating the action of 1-amino-3-(N,N-methylamino)-propylidene-1,1-bisphosphonic acid, which can be blocked by Ca2+-channel blockers it is preferred that the Ca2+-channel blockers are selected from the group comprising nifedipine and verapamil.
In one embodiment it is preferred that 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid be used in doses of 0.01 to 1000 mg/oral application, more preferably 12.5 to 75 mg/oral application. The term xe2x80x9coral applicationxe2x80x9d is meant to include solid or soluble liquid pharmaceutical formulations, gels, soft capsules, tablets, capsules containing solid preparations, soluble liquid forms or suspensions, and pills. Within one aspect of the present invention it is envisioned that 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid be used also for topical administration to be applied on skin and/or mucosae. Forms of application useful for this purpose comprise ointments, creams, sprays, suppositories and gels.
In another embodiment of the present invention it is preferred that 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid be used in doses of 0.02 to 200 mg/parenteral application, more preferably 2.5 to 15 mg/parenteral application. The term xe2x80x9cparenteral applicationxe2x80x9d is meant to include any application that avoids the gastro-intestinal tract, for example by sub-cutaneous, intra-muscular or intra-venous injection or infusion.
The term xe2x80x9cIP3-sensitive storesxe2x80x9d refers to those intracellular stores of Ca2+-ions which, upon the presence of IP3 (=inositol triphosphate) rapidly release Ca2+-ions. Such IP3-sensitive stores encompass for example the endoplasmic reticulum and, in smooth muscle cells, the sarcoplasmic reticulum.
It is to be noted that according to the present invention the influence exerted by 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid on the Ca2+ homeostasis of the osteoblasts, in particular an increase of the Ca2+-levels in the osteoblasts, can occur in at least two different ways: Ca2+-ions can be mobilized from endogenous stores, such as those which are IP3-sensitive or thapsigargin-senstitive.
Secondly, there can also occur a Ca2+-ion-influx from extracellular stores. This may for example occur through voltage-dependent Ca2+-channels (VDCC), but other forms of Ca2+-channels, e.g. ligand-gated channels, can be envisioned, too.
It is within the scope of the present invention that 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid also be used for screening for Ca2+-channel blockers. The term xe2x80x9cputative Ca2+-channel blockerxe2x80x9d is meant to designate a compound to be tested for its Ca2+-channel-blocking capabilities.
The term xe2x80x9cputative functional analoguexe2x80x9d of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid designates any compound that is known to cause a Ca2+-ion influx into the cells in the absence of any Ca2+-channel blockers and is therefore mimicking 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid in its influence on Ca2+-homeostasis of the cells. The term xe2x80x9cmeasuring a responsexe2x80x9d is also meant to include the measurement or detection of the absence of a response. Within the present invention, the term xe2x80x9cresponsexe2x80x9d is preferably meant to designate a change in cytosolic Ca2+-levels.
It is also within the scope of the present invention to combine 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid in its novel uses together with at least one other compound/substance, in synergistic combinations. The term xe2x80x9ccombinationxe2x80x9d is meant to include the incorporation of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid together with another (other) substance(s) into the same medicament, as well as the administering of a medicament comprising 1-amino-3-(N,N-diethylamino)-propylidene-1,1-bisphosphonic acid to a patient with another (other) substance(s), i.e. administering this (these) other substance(s) in a physically separated, i.e. spatially or temporally separated manner from the medicament. In the case where the substance(s) is (are) administered in a separated manner from the medicament comprising 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid, it is preferred that the 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid-comprising medicament is administered with the other substance(s) in a single-step (concerted intake, infusions, injections), sequential-step (one after the other) or cyclic-step fashion. In one embodiment it is preferred that the medicament comprising 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid is applied before, during or after treatment with other amino-substituted bisphosphonates. For example, a sequential administering of the 1-amino-3-(N,N-dimethylamino)-propylidene-1,-bisphosphonic acid-comprising medicament with other amino-substituted bisphosphonates may be indicated, since other amino-substituted bisphosphonates exert a partial anti-resorptive effect. So a patient may need a 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid-comprising medicament for bone formation due to the selective modulating effect of 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid on the osteoblasts, and at the same time another amino-substituted bisphosphonate for a xe2x80x9ctunablexe2x80x9d degree of inhibition of bone resorption, depending on his/her individual requirement.
As another example a patient might first require a high degree of inhibition of bone resorption, continue then with low or none and then change again afterwards, according to individual requirements which can be determined by biochemical, densitometrical, X-ray or other measurements performed on the bones of the individual.
Thus 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid is an ideal candidate forming the basis for an individually tunable therapy of conditions where bones are weak and/or fragile.
This substance/these substances/compounds of the aforementioned synergistic combinations is/are selected from the group comprising calcium salts, calcium citrate and calcium carbonate, other amino-substituted bisphosphonates, pharmaceutically active fluorine-containing salts, vitamins of the D-group and their metabolites, cholecalciferol, calcifediol, calcitriol, ergocalciferol, PTH, anabolic hormones, such as estrogens, substances with estrogenic activity on the bone, progestogens, androgens, growth hormones, peptides with growth hormone activity, selective modulators of the estrogenic receptor, raloxifene and others. The term xe2x80x9camino-substituted bisphosphonatexe2x80x9d preferably designates a bisphosphonate compound that has an amino group at the 1-position. Other substitution positions, however, can be envisioned, too. Where the xe2x80x9caminoderivativexe2x80x9d of a 1-hydroxy bisphosphonate is mentioned here, this is meant to encompass molecules having a structure in which the hydroxy-group is replaced by an amino group.
It is one of the surprising advantages of the present invention that 1-amino-3-(N,N-dimethylamino)-propylidene-1,1-bisphosphonic acid acts to avoid the formation of the aforementioned xe2x80x9cunfitxe2x80x9d bone structures and aids in removing them once they have been formed due to its selective modulating capabilities with respect to the osteoblasts.
Therefore, unfit mineralized structures in the bone are prevented and removed respectively. For example, these structures can arise in older people, but have not manifested themselves in clinically pathological conditions yet. Another important example where these unfit structures may occur is the growing skeleton, i.e. children. These may be healthy children or children having an osteopathy. The present invention provides means for removing these unfit structures or preventing them in the first place, thus contributing to a healthy bone structure which is capable of withstanding the mechanical stress exerted upon the bone through daily usage.
These and other aspects of the present invention will become evident upon reference to the following examples and attached figures. In addition, various references are set forth below which describe in more detail certain procedures or experimental details and are therefore incorporated by reference in their entirety.